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Team:Todai-Tokyo/Notebook/bread
From 2009.igem.org
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*TA cloning of mtlD<BR> | *TA cloning of mtlD<BR> | ||
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*Miniprep of mtlD<BR> | *Miniprep of mtlD<BR> | ||
Revision as of 03:36, 16 October 2009
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Contents |
Plan
Aim:Create yeast that can be used to make sweet and low energy bread
Methods:
- Clone the glucoamylase gene from Saccharomycopsis fibuligera and insert it in the yeast chromosome by homologous recombination
- Clone mtlD (mannitol synthase) from E.coli and insert it in the yeast chromosome by homologous recombination
- Replace gpd1 gene by Glu1 and gpd2 gene by mtlD
~9/20
- PCR of mtlD
- PCR of gpd1 promoter
- TA cloning of mtlD
9/21
- Miniprep of mtlD
9/22
- read the sequence of mtlD→successful
- mtlD primers with HAtag come
9/25
- PCR of mtlD with new primer→failed
9/26
- PCR of mtlD with new primer→successful
9/27
- PCR of gpd1 promoter with Pfu Ultra
October
- PCR of Glu1
- TA cloning of Glu1
- PCR of gpd1 promoter with ExTaq
10/14
- cut mtlD and plate1 7D both by EcoRI and PstI
- colony PCR of Glu1
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